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== Ethanol precipitation == * Pool 20 reactions into 2x 15 mL tube * Add 2.5x 100% ethanol, 0.1x 3M sodium acetate, and 8 uL of glycoblue * Store in -80C for at least 1 hour. * Spin at >4000 rpm for 40 minutes at 4C * Remove supernatant & add 1 mL 80% ice-cold ethanol * Spin at >4000 rpm for 15 minutes at 4C * Remove supernatant & air dry for 10 minutes * Add 150 uL of water to each tube & pool together. * Purify with 3x Qiaquick PCR cleanup columns. (Optional: dry residual ethanol by warming columns in thermomixer for 5 minutes at 50C). Elute each with 30 uL of pre-warmed EB. * Nanodrop to quantify yield.
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